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Proceedings Paper

Photo-bleaching compensation for autofocus algorithms in fluorescence microscope applications
Author(s): Paolo Soda; Giulio Iannello
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Paper Abstract

In this paper, we report our experience in looking for effective autofocus functions for automatic image acquisition in ImmunoFluorescence Assay (IFA). We propose to use two functions that greatly improve the performance with respect to functions commonly proposed in the literature. The first function is based on the image histogram and it is suited to in the coarse phase of autofocus, when the z-axis steps are larger to speedup the identification of the interval where lies the focus position. The second function is a popular autofocus function properly modified to compensate the effects of photo-bleaching. It is best suited to be used in the other phases of the autofocus process when smaller steps are taken to precisely identify the focus position. Effectiveness of the proposed functions has been assessed on real images both quantitatively and qualitatively, confirming that they allow obtaining a high quality images, in most cases better than those manually acquired.

Paper Details

Date Published: 2 March 2006
PDF: 7 pages
Proc. SPIE 6142, Medical Imaging 2006: Physics of Medical Imaging, 61423S (2 March 2006); doi: 10.1117/12.650096
Show Author Affiliations
Paolo Soda, Univ. Campus Bio-Medico di Roma (Italy)
Giulio Iannello, Univ. Campus Bio-Medico di Roma (Italy)

Published in SPIE Proceedings Vol. 6142:
Medical Imaging 2006: Physics of Medical Imaging
Michael J. Flynn; Jiang Hsieh, Editor(s)

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